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Optimization of Culture Conditions for Production of the Anti-Leukemic Glutaminase Free L-Asparaginase by Newly Isolated Streptomyces olivaceus NEAE-119 Using Response Surface Methodology.

El-Naggar Nel-A, Moawad H, El-Shweihy NM, El-Ewasy SM - Biomed Res Int (2015)

Bottom Line: The optimization of different process parameters for L-asparaginase production by Streptomyces olivaceus NEAE-119 using Plackett-Burman experimental design and response surface methodology was carried out.Fifteen variables (temperature, pH, incubation time, inoculum size, inoculum age, agitation speed, dextrose, starch, L-asparagine, KNO3, yeast extract, K2HPO4, MgSO4·7H2O, NaCl, and FeSO4·7H2O) were screened using Plackett-Burman experimental design.The most positive significant independent variables affecting enzyme production (temperature, inoculum age, and agitation speed) were further optimized by the face-centered central composite design-response surface methodology.

View Article: PubMed Central - PubMed

Affiliation: Department of Bioprocess Development, Genetic Engineering and Biotechnology Research Institute, City of Scientific Research and Technological Applications, Alexandria 21934, Egypt.

ABSTRACT
Among the antitumor drugs, bacterial enzyme L-asparaginase has been employed as the most effective chemotherapeutic agent in pediatric oncotherapy especially for acute lymphoblastic leukemia. Glutaminase free L-asparaginase producing actinomycetes were isolated from soil samples collected from Egypt. Among them, a potential culture, strain NEAE-119, was selected and identified on the basis of morphological, cultural, physiological, and biochemical properties together with 16S rRNA sequence as Streptomyces olivaceus NEAE-119 and sequencing product (1509 bp) was deposited in the GenBank database under accession number KJ200342. The optimization of different process parameters for L-asparaginase production by Streptomyces olivaceus NEAE-119 using Plackett-Burman experimental design and response surface methodology was carried out. Fifteen variables (temperature, pH, incubation time, inoculum size, inoculum age, agitation speed, dextrose, starch, L-asparagine, KNO3, yeast extract, K2HPO4, MgSO4·7H2O, NaCl, and FeSO4·7H2O) were screened using Plackett-Burman experimental design. The most positive significant independent variables affecting enzyme production (temperature, inoculum age, and agitation speed) were further optimized by the face-centered central composite design-response surface methodology.

No MeSH data available.


Related in: MedlinePlus

Scanning electron micrograph showing the spore chain morphology and spore surface ornamentation of strain NEAE-119 grown on starch nitrate agar medium for 14 days at 30°C at magnification of 3000x (a), 15000x (b), and 22000x (c).
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fig2: Scanning electron micrograph showing the spore chain morphology and spore surface ornamentation of strain NEAE-119 grown on starch nitrate agar medium for 14 days at 30°C at magnification of 3000x (a), 15000x (b), and 22000x (c).

Mentions: Morphological observation of the 14-day-old culture of strain NEAE-119 grown on yeast extract-malt extract agar (ISP 2) [28] revealed that strain NEAE-119 had the typical characteristics of the genus Streptomyces [40]; it is aerobic and mesophilic; both aerial and vegetative hyphae were abundant, well-developed, and not fragmented. Aerial mycelium color was varied from the grey color to greyish beige or whitish grey on different test media. Cultural characteristics of strain NEAE-119 are shown in the table in the Supplementary Materials available online at http://dx.doi.org/10.1155/2015/627031. Strain NEAE-119 grew well on yeast extract-malt extract agar (ISP medium 2), oatmeal agar (ISP medium 3), inorganic salt-starch agar (ISP medium 4), glycerol-asparagine agar (ISP medium 5), peptone-yeast extract iron agar (ISP medium 6), and tyrosine agar (ISP medium 7). Verticils are not present. The mycelium does not fragment. It formed an extensively branched substrate mycelium and aerial hyphae which differentiated into spore chains. Spore chains with many spores were in section Spirals, with open spirals intergrading through flexuous spore chains suggestive of section Rectiflexibiles. Mature spore chains are generally long, often with more than 50 spores per chain. This morphology is seen on starch nitrate agar medium. Spore surface is smooth (0.55–0.90 × 1.16–1.34 μm in diameter) (Figure 2).


Optimization of Culture Conditions for Production of the Anti-Leukemic Glutaminase Free L-Asparaginase by Newly Isolated Streptomyces olivaceus NEAE-119 Using Response Surface Methodology.

El-Naggar Nel-A, Moawad H, El-Shweihy NM, El-Ewasy SM - Biomed Res Int (2015)

Scanning electron micrograph showing the spore chain morphology and spore surface ornamentation of strain NEAE-119 grown on starch nitrate agar medium for 14 days at 30°C at magnification of 3000x (a), 15000x (b), and 22000x (c).
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4477217&req=5

fig2: Scanning electron micrograph showing the spore chain morphology and spore surface ornamentation of strain NEAE-119 grown on starch nitrate agar medium for 14 days at 30°C at magnification of 3000x (a), 15000x (b), and 22000x (c).
Mentions: Morphological observation of the 14-day-old culture of strain NEAE-119 grown on yeast extract-malt extract agar (ISP 2) [28] revealed that strain NEAE-119 had the typical characteristics of the genus Streptomyces [40]; it is aerobic and mesophilic; both aerial and vegetative hyphae were abundant, well-developed, and not fragmented. Aerial mycelium color was varied from the grey color to greyish beige or whitish grey on different test media. Cultural characteristics of strain NEAE-119 are shown in the table in the Supplementary Materials available online at http://dx.doi.org/10.1155/2015/627031. Strain NEAE-119 grew well on yeast extract-malt extract agar (ISP medium 2), oatmeal agar (ISP medium 3), inorganic salt-starch agar (ISP medium 4), glycerol-asparagine agar (ISP medium 5), peptone-yeast extract iron agar (ISP medium 6), and tyrosine agar (ISP medium 7). Verticils are not present. The mycelium does not fragment. It formed an extensively branched substrate mycelium and aerial hyphae which differentiated into spore chains. Spore chains with many spores were in section Spirals, with open spirals intergrading through flexuous spore chains suggestive of section Rectiflexibiles. Mature spore chains are generally long, often with more than 50 spores per chain. This morphology is seen on starch nitrate agar medium. Spore surface is smooth (0.55–0.90 × 1.16–1.34 μm in diameter) (Figure 2).

Bottom Line: The optimization of different process parameters for L-asparaginase production by Streptomyces olivaceus NEAE-119 using Plackett-Burman experimental design and response surface methodology was carried out.Fifteen variables (temperature, pH, incubation time, inoculum size, inoculum age, agitation speed, dextrose, starch, L-asparagine, KNO3, yeast extract, K2HPO4, MgSO4·7H2O, NaCl, and FeSO4·7H2O) were screened using Plackett-Burman experimental design.The most positive significant independent variables affecting enzyme production (temperature, inoculum age, and agitation speed) were further optimized by the face-centered central composite design-response surface methodology.

View Article: PubMed Central - PubMed

Affiliation: Department of Bioprocess Development, Genetic Engineering and Biotechnology Research Institute, City of Scientific Research and Technological Applications, Alexandria 21934, Egypt.

ABSTRACT
Among the antitumor drugs, bacterial enzyme L-asparaginase has been employed as the most effective chemotherapeutic agent in pediatric oncotherapy especially for acute lymphoblastic leukemia. Glutaminase free L-asparaginase producing actinomycetes were isolated from soil samples collected from Egypt. Among them, a potential culture, strain NEAE-119, was selected and identified on the basis of morphological, cultural, physiological, and biochemical properties together with 16S rRNA sequence as Streptomyces olivaceus NEAE-119 and sequencing product (1509 bp) was deposited in the GenBank database under accession number KJ200342. The optimization of different process parameters for L-asparaginase production by Streptomyces olivaceus NEAE-119 using Plackett-Burman experimental design and response surface methodology was carried out. Fifteen variables (temperature, pH, incubation time, inoculum size, inoculum age, agitation speed, dextrose, starch, L-asparagine, KNO3, yeast extract, K2HPO4, MgSO4·7H2O, NaCl, and FeSO4·7H2O) were screened using Plackett-Burman experimental design. The most positive significant independent variables affecting enzyme production (temperature, inoculum age, and agitation speed) were further optimized by the face-centered central composite design-response surface methodology.

No MeSH data available.


Related in: MedlinePlus